title: Structural basis for the different stability and activity between the Cdk5 complexes with p35 and p39 activators.
abstract: Cyclin-dependent kinase 5 (Cdk5) is a brain-specific membrane-bound protein kinase that is activated by binding to the p35 or p39 activator. Previous studies have focused on p35-Cdk5, and little is known regarding p39-Cdk5. The lack of functional understanding of p39-Cdk5 is due, in part, to the labile property of p39-Cdk5, which dissociates and loses kinase activity in nonionic detergent conditions. Here we investigated the structural basis for the instability of p39-Cdk5. p39 and p35 contain N-terminal p10 regions and C-terminal Cdk5 activation domains (AD). Although p35 and p39 show higher homology in the C-terminal AD than the N-terminal region, the difference in stability is derived from the C-terminal AD. Based on the crystal structures of the p25 (p35 C-terminal region including AD)-Cdk5 complex, we simulated the three-dimensional structure of the p39 AD-Cdk5 complex and found differences in the hydrogen bond network between Cdk5 and its activators. Three amino acids of p35, Asp-259, Asn-266, and Ser-270, which are involved in hydrogen bond formation with Cdk5, are changed to Gln, Gln, and Pro in p39. Because these three amino acids in p39 do not participate in hydrogen bond formation, we predicted that the number of hydrogen bonds between p39 and Cdk5 was reduced compared with p35 and Cdk5. Using substitution mutants, we experimentally validated that the difference in the hydrogen bond network contributes to the different properties between Cdk5 and its activators.
authors.affiliation: From the Department of Biological Sciences, Tokyo Metropolitan University, Minami-osawa, Hachioji, Tokyo 192-0397.
authors.forename: Taro
authors.initials: T
authors.lastname: Saito
authors.affiliation: From the Department of Biological Sciences, Tokyo Metropolitan University, Minami-osawa, Hachioji, Tokyo 192-0397.
authors.forename: Masashi
authors.initials: M
authors.lastname: Yano
authors.affiliation: From the Department of Biological Sciences, Tokyo Metropolitan University, Minami-osawa, Hachioji, Tokyo 192-0397.
authors.forename: Yusei
authors.initials: Y
authors.lastname: Kawai
authors.affiliation: From the Department of Biological Sciences, Tokyo Metropolitan University, Minami-osawa, Hachioji, Tokyo 192-0397.
authors.forename: Akiko
authors.initials: A
authors.lastname: Asada
authors.affiliation: Celestar Lexico-Sciences, Inc., Nakase, Mihama-ku, Chiba 261-8501.
authors.forename: Mitsuhito
authors.initials: M
authors.lastname: Wada
authors.affiliation: Celestar Lexico-Sciences, Inc., Nakase, Mihama-ku, Chiba 261-8501; the Research Center for Advanced Science and Technology, The University of Tokyo, Komaba, Meguro, Tokyo 153-8914, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Doi
authors.affiliation: From the Department of Biological Sciences, Tokyo Metropolitan University, Minami-osawa, Hachioji, Tokyo 192-0397,. Electronic address: hisanaga-shinichi@tmu.ac.jp.
authors.forename: Shin-Ichi
authors.initials: SI
authors.lastname: Hisanaga
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051360:Cyclin-Dependent Kinase 5 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003598:Cytoskeletal Proteins; D057809:HEK293 Cells; D006801:Humans; D006860:Hydrogen Bonding; D058635:Lipid-Linked Proteins; D051379:Mice; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020125:Mutation, Missense; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D055550:Protein Stability; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship
chemical_list: D002352:Carrier Proteins; C578513:Cdk5r2 protein, mouse; D003598:Cytoskeletal Proteins; D058635:Lipid-Linked Proteins; D046912:Multiprotein Complexes; D009419:Nerve Tissue Proteins; C089495:neuronal Cdk5 activator (p25-p35); D051360:Cyclin-Dependent Kinase 5; C495905:CDK5 protein, human; C495906:Cdk5 protein, mouse
keywords: CDK (Cyclin-dependent Kinase); Cdk5; Computer Modeling; Cyclins; Hydrogen Bond; Neurons; Serine-Threonine Protein Kinase; Subunit Interaction; p35; p39